Objective To investigate the setup error in patients with spinal bone metastasis who underwent radiotherapy under the guidance of kilovoltage cone-beam CT (KV-CBCT). Methods A total of 118 patients with spinal metastasis who underwent radiotherapy, including 17 cases of cervical spine, 62 cases of thoracic spine, and 39 cases of lumbar spine, were collected. KV-CBCT scans were performed using the linear accelerators from Elekta and Varian’s EDGE system. CBCT images were registered with reference CT images in the bone window mode. A total of 973 data were collected, and 3D linear errors were recorded. Results The patients with spinal bone metastasis were grouped by site, height, weight, and BMI. The P value of the patients grouped only by site was P<0.05, which was statistically significant. Conclusion When grouped by site in the 3D direction, the positioning effect of cervical spine is better than that of thoracic and lumbar spine. The positioning effect of the thoracic spine is better in the head and foot direction but worse in the left and right direction compared with that of the lumbar spine. Instead of extending or narrowing the margin according to the BMI of patients with spinal metastasis, the margin must be changed according to the site of spinal bone metastasis.

Alzheimer's disease (AD) is the major form of dementia in the elderly and is closely related to the toxic effects of microglia sustained activation. In AD, sustained microglial activation triggers impaired synaptic pruning, neuroinflammation, neurotoxicity, and cognitive deficits. Accumulating evidence has demonstrated that aberrant expression of deubiquitinating enzymes is associated with regulating microglia function. Here, we use RNA sequencing to identify a deubiquitinase YOD1 as a regulator of microglial function and AD pathology. Further study showed that YOD1 knockout significantly improved the migration, phagocytosis, and inflammatory response of microglia, thereby improving the cognitive impairment of AD model mice. Through LC-MS/MS analysis combined with Co-IP, we found that Myosin heavy chain 9 (MYH9), a key regulator maintaining microglia homeostasis, is an interacting protein of YOD1. Mechanistically, YOD1 binds to MYH9 and maintains its stability by removing the K48 ubiquitin chain from MYH9, thereby mediating the microglia polarization signaling pathway to mediate microglia homeostasis. Taken together, our study reveals a specific role of microglial YOD1 in mediating microglia homeostasis and AD pathology, which provides a potential strategy for targeting microglia to treat AD.

Platelet transfusion is one of the critical clinical therapies of neonatal thrombocytopenia and significant preventive measures of bleeding diseases in preterm infants.The platelet transfusion rate is high in clinical practice，whereas some controversies emerge in its clinical application.Platelet transfusion decision-making should take into account the platelet count and the causes of thrombocytopenia.In addition，the presence of bleeding tendency and platelet effects on other systemic disorders should be considered.Clinicians often need to make rapid decisions about whether to transfuse platelets in a critically situation.On the basis of the comprehensive overview of issues that are closely pertinent to the field of clinical practice，this article is dedicated to elucidate the advancements in clinical research pertaining to neonatal platelet transfusions，aiming to serve as a reference for clinicians when making transfusion decisions and to chart a course for future clinical investigations.

Aim To explore the effect of succinate/G protein coupled receptor 91(GPR91)on mitochondria in vascular endothelial cells and its regulatory mechanisms.Methods Transmission electron microscopy,Western blot and fluorescence microscopy were used to observe the effects of succinate analogues diethyl succinate(DS),GPR91 agonist and inhibitor on the mitochondrial morphology,cristae,cristate homeostasis related proteins reactive oxygen species(ROS)content,Ca2+concentration,mitochondrial membrane potential,the expression of dihydroorotate dehydrogenase(DHODH)and oxidized coenzyme Q10(CoQlO).Fluorescence probes were used to observe the effect of DHODH inhib-itor and CoQ10 on ROS level and Ca2+concentration of endothelial cells.Results After DS treatment,the mitochon-dria showed pyknosis and mitochondrial volume significantly decreased,electron density of the mitochondrial membrane in-creased,and the number of cristae decreased in endothelial cells;the expression of cristae homeostasis related proteins MIC60 decreased by 23％,while cellular ROS level and Ca2+concentration increased;mitochondrial membrane potential decreased(P＜0.05 or P＜0.01).After GPR91 agonist treatment,the expression of cristae homeostasis related proteins MIC60 decreased by 31％,meanwhile,cellular ROS level increased by 27％and Ca2+concentration increased by 36％;mitochondrial membrane potential decreased(P＜0.05 or P＜0.01).After GPR91 inhibitor treatment,the expression of cristae homeostasis related proteins MIC60 increased by 22％and ATP5I increased by 40％;the levels of ROS decreased by 41％and Ca2+concentration decreased by 67％;and the mitochondrial membrane potential was restored to normal(P＜0.05 or P＜0.01).After DS treatment,the expression of DHODH decreased by 43％and the level of oxidized CoQ10 in-creased by 120％(P＜0.05 or P＜0.01).After GPR91 agonist treatment,the expression of DHODH decreased by 22％and the level of oxidized CoQ10 increased by 36％(P＜0.05 or P＜0.01).After GPR91 inhibitor treatment,the expres-sion of DHODH increased by 40％and the level of oxidized CoQ10 decreased by 39％(P＜0.01).After DHODH inhibi-tor treatment,the ROS level increased by 20％and Ca2+concentration increased by 28％,and mitochondrial membrane po-tential reduced at same time(P＜0.05 or P＜0.01).Exogenous oxidized CoQ10 inhibited ROS production by 30％and decreased Ca2+concentration by 20％(P＜0.05 or P＜0.01).Conclusion Succinate/GPR91 promotes mitochondrial damage in endothelial cells,and its mechanism may relate to down-regulating the expression of DHODH and inhibiting the reduction of CoQ10 by affecting the mitochondrial cristae homeostasis.

Objective:To analyze the time needed for active breathing coordinator (ABC) coaching in tumor patients, and to explore the influencing factors of coaching time.Methods:A retrospective study was conducted on 93 patients who received ABC treatment led by the same staff at the Cancer Hospital of Chinese Academy of Medical Sciences from September 2019 to April 2021. The effects of education level, body mass index (BMI), age, gender and disease type on the couching time were analyzed. The coaching time was expressed as Mean ± SD. Independent sample t-test or rank sum test was used for comparison between different groups. P<0.05 was considered statistically significant. Results:Statistical significance was observed in the effect of education level, BMI and age on coaching time. The coaching time in the higher education group was (9.74±3.80) min, significantly shorter than the (13.79±6.03) min ( P=0.001) of the primary education group and the (13.03±5.14) min ( P=0.021) of the middle education group. The couching time in the BMI<24 kg/m 2 group was (10.27±3.98) min, significantly shorter compared with (12.74±5.60) min ( P<0.001) in the BMI≥24 kg/m 2 group. The coaching time in the ≥60 years old group was (14.12±5.06) min, significantly longer than the (9.86±3.76) min ( P=0.002) of the ≤40 years old group and the (11.30±5.10) min ( P=0.021) of the 40-60 years old group. No significant differences were noted in the effect of gender, disease type and tumor staging on the coaching time. The coaching time in males and females was (13.54±5.89) and (10.94±4.61) min, respectively ( P=0.071). The coaching time of patients with breast cancer, lung cancer, liver cancer, mediastinal lymphoma and pancreatic cancer was (10.75±4.72), (15.30±5.57), (11.69±4.96), (9.86±3.61) and (12.15±0.07) min, respectively ( P=0.071). The coaching time of stageⅠ,Ⅱ,Ⅲ and Ⅳ patients was (10.35±4.37), (11.88±5.30), (9.52±2.51) and (14.32±5.27) min ( P=0.060). Conclusions:Patients with higher education level and BMI<24 kg/m 2 require less ABC coaching time. Patients aged≥60 years require longer coaching time. Gender, disease type and clinical stage exert no significant effect on the duration of coaching.

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.

Objective To investigate the endocytosis and exocytosis of soluble uranium in human kidney proximal tubular epithelial (HK-2) cells and the cytotoxicity after uranium exposure. Methods Cell Counting Kit-8 assay was used to determine the cell viability after different concentrations of uranium exposure, and optical microscopy and transmission electron microscopy were used to observe the changes in cells after uranium exposure. Inductively coupled plasma mass spectrometry was used to monitor the endocytosis and exocytosis of uranium over time by cells. Flow cytometry was used to assess the changes in cell cycle and apoptosis after uranium exposure. Results After uranium exposure, HK-2 cells showed dose-dependent damage; cell cycle was arrested in G1 phase; cell apoptosis and necrosis occurred; cell proliferation was inhibited. The content of endocytic uranium increased gradually within 24 h, and there was a threshold for uranium endocytosis, while the fraction of uranium binding to cell surface was low (< 0.2%). Over 40% of the endocytic uranium would be exocytosed within 1 h. Uranium could form needle-like precipitates in both intracellular and extracellular areas after uranium exposure. Conclusion After uranium exposure, cells show decreased viability, cell cycle arrest, and cell apoptosis. The process of endocytosis and exocytosis of soluble uranium is very rapid. HK-2 cells can convert soluble uranium into non-toxic precipitates.

Objective:To select and optimize the conditions of urine cobalt determination by graphite furnace atomic absorption spectroscopy and establish a method for urine cobalt determination.Methods:In April 2020, the matrix modifier and spectrum wavelength were selected by mathematical statistics method, the heating procedure of graphite furnace was optimized, the variance of orthogonal test results was analyzed, and the working curve was quantified by external standard method.Results:The results showed that the diammonium hydrogen phosphate was a matrix modifier with a wavelength of 240.7 nm, and the optimized graphite furnace heating procedure showed a good linear relationship at 5-80 μg/L. The correlation coefficient was 0.9991. The detection limit was 0.9 μg/L, the lowest detection concentration was 1.8 μg/L. The recovery was 94.0%-101.4%, and the precision was 2.6%-5.8%.Conclusion:The orthogonal design method can efficiently and scientifically optimize the optimal combination of the various factor levels of graphite furnace atomic absorption spectrometry. The determination method established based on this has high precision and good accuracy, and can be applied to the determination of cobalt in urine.

Objective:To select and optimize the conditions of urine cobalt determination by graphite furnace atomic absorption spectroscopy and establish a method for urine cobalt determination.Methods:In April 2020, the matrix modifier and spectrum wavelength were selected by mathematical statistics method, the heating procedure of graphite furnace was optimized, the variance of orthogonal test results was analyzed, and the working curve was quantified by external standard method.Results:The results showed that the diammonium hydrogen phosphate was a matrix modifier with a wavelength of 240.7 nm, and the optimized graphite furnace heating procedure showed a good linear relationship at 5-80 μg/L. The correlation coefficient was 0.9991. The detection limit was 0.9 μg/L, the lowest detection concentration was 1.8 μg/L. The recovery was 94.0%-101.4%, and the precision was 2.6%-5.8%.Conclusion:The orthogonal design method can efficiently and scientifically optimize the optimal combination of the various factor levels of graphite furnace atomic absorption spectrometry. The determination method established based on this has high precision and good accuracy, and can be applied to the determination of cobalt in urine.

Objective: To evaluate the long-term ef efficacy and safety of topical 1% atropine for retarding pregressive myopia. Methods: A randomized controlled study evaluating atropine and placebo in 570 Chinese children aged 8~14 years recruited from pediatric ophthalmology in Yunnan Provincial the Second People s Hospital during Jan. 2015 to Dec. 2019. In experimental group, patients received drops every two weeks for 24 months, then every three weeks for 12 months, followed by no drops for 12 months. In control group, all children wear single focus frame glasses. Spherical equivalent, axial length, intraocular pressure and atropinerelated side effects were examined at 6, 12, 24, 36 and 48 months for all children. Results: At the end of stage Ⅰ, the myopia progression in the atropine treatment group (-0.27±0.81)D was significantly lower than that in the control group (-1.29±0.13)D, and the increase of axial length in the atropine group (0.11±0.13)mm was also significantly lower than that in the control group (0.41±0.19)mm (P<0.05). At the end of stage Ⅱ, the average myopia progression in the atropine treatment group (-0.31±0.28)D was significantly lower than that in the control group (-0.80±0.66)D (P<0.01). Similarly, the axial growth of the experimental group (0.14±0.09)mm was significantly lower than that of the control group (0.39±0.14)mm (P<0.01). After the withdrawal of atropine eye drops (stage Ⅲ), there was no significant refractive regression in the experimental group. During the whole follow-up period, no serious adverse events related to atropine were found. Conclusion Local intermittent use of 1% atropine eye drops and the gradual reduction of atropine eye drops can ensure the effectiveness in the treatment of myopia, reducing the side effects of atropine, avoiding refractive regression after drug withdrawal, and improving children s compliance at the same time.